The Rat Melatonin ELISA Kit is specifically designed for the accurate detection of melatonin levels in rat serum, plasma, and tissue culture supernatants. With its high sensitivity and specificity, this kit delivers dependable and consistent results, making it a valuable tool for research in various fields. Melatonin is a hormone that regulates the sleep-wake cycle and is involved in numerous physiological processes, including immune function, antioxidant activity, and reproductive health. Dysregulation of melatonin levels has been linked to various health conditions, such as insomnia, depression, and metabolic disorders, highlighting its importance as a biomarker for studying these conditions and exploring potential treatments.
With easy-to-follow protocols and quick assay turnaround times, the Rat Melatonin ELISA Kit offers convenience and flexibility for researchers looking to investigate the role of melatonin in health and disease. Order yours today and unlock new insights into the fascinating world of melatonin biology.
Product Name:
Rat Melatonin ELISA Kit
SKU:
RTFI00972
Reactivity:
Rat
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
4.688 pg/mL
Range:
7.813-500 pg/mL
Sample Type:
Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples
Storage:
2-8°C(Sealed), Don't cryopreserve.
Linearity:
Sample
1:2
1:4
1:8
Serum (n = 10)
87-104%
86-99%
81-99%
EDTA Plasma (n = 10)
89-103%
85-101%
84-100%
Heparin Plasma (n = 10)
90-103%
87-101%
84-98%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
86-105
96
EDTA Plasma (n = 10)
85-103
94
Heparin Plasma (n = 10)
89-104
96
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Procedure
1
Reagent & Plate Preparation: Equilibrate TMB substrate for 30 minutes at room temperature. Prepare standards, samples (minimum 1:2 dilution), blanks, assign wells, and pre-wash the plate twice.
2
Sample & Biotin-Antibody Binding: Add 50 µL standard or sample followed by 50 µL biotin-labeled antibody to each well. Mix gently and incubate at 37°C for 45 minutes.
3
Washing: Wash the plate 5 times with wash buffer, allowing 1 minute soak time per wash.
4
Color Development: Add TMB substrate and incubate in the dark at 37°C for 10-20 minutes until color develops.
5
Stop Reaction: Add stop solution to terminate the reaction. The color changes from blue to yellow immediately.
6
Reading: Measure absorbance at 450 nm using a microplate reader.
Sample Type
Protocol
Serum
Allow blood to clot, centrifuge at 1000 × g for 20 minutes, collect supernatant and store appropriately.
Plasma
Collect using EDTA anticoagulant, centrifuge at 1000 × g for 15 minutes at 2–8°C and collect plasma.
Cell Culture Supernatant
Centrifuge at 1000 × g for 20 minutes at 4°C and collect clarified supernatant.
Cell Lysate
Lyse cells using recommended lysis buffer with protease inhibitors, centrifuge at 10,000 rpm for 10 minutes, and collect protein supernatant.
Tissue Homogenate
Homogenize tissue in PBS with protease inhibitors, centrifuge at 5000 × g for 5 minutes, and collect supernatant.
Other Sample Types
Centrifuge samples at 1000 × g for 15 minutes at 2–8°C and collect supernatant. For additional guidance, please contact techsupport@assaygenie.com.
Component
Quantity
Storage
48T
96T
ELISA Microplate (Dismountable)
8×6
8×12
Place the test strips into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
Lyophilized Standard
1 vial
2 vial
Place the standards into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
Biotin-labeled Antibody (Concentrated, 100X)
60 ul
120 ul
2-8°C (Avoid direct light)
HRP-Streptavidin Conjugate (SABC, 100X)
60 ul
120 ul
2-8°C (Avoid direct light)
TMB Substrate
5 ml
10 ml
2-8°C (Avoid direct light)
Sample Dilution Buffer
10 ml
20 ml
2-8°C
Antibody Dilution Buffer
5 ml
10 ml
2-8°C
SABC Dilution Buffer
5 ml
10 ml
2-8°C
Stop Solution
5 ml
5 ml
2-8°C
Wash Buffer(25X)
15 ml
30 ml
2-8°C
Plate Sealer
3 pieces
5 pieces
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Technical Manual
1 copy
1 copy
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Alexis Makenna Carmine et al.
Ultrasound Modulation of Melatonin Release from Pineal Glands in Vitro
